Objective To analyze the differences in protein pro fi le between laterally spreading tumor (LST) cell line and SW480 cell line using t wo-dimensional electrophoresis (2-DE). Methods 2-DE was empl oyed to isolate the total proteins of the two cell lines.The gels were stained with silver, and the differential protein expressions were analyzed with Melanin e 3 software. Results The protein spots of the two cell lines w ere 1285?51(LST) and 1184?47(SW480) respectively,containing 96?7 differential protein spots with 50?6 expressed or obviously increased only in LST cell line and 47?5 in SW480 cell line. Conclusion The protein profiles between LST and SW480 cell lins are different and further analysis on the differ ent interested spots is required to acquire the biology-associated proteins spe cific LST.

Objective To observe the influence on adhesion between LST-R1 cell and collagen Ⅰ mediated by galectin-1 antibody inhibition. Methods Laser confocal scanning microscopy was employed to detect the expression of galectin-1 on LST-R1 cell membrane.LST-R1 cells inhibited by galectin-1 antibody or not were seeded in the 48-well plate coated with collagen Ⅰ (20?g/well), and the morphology of the adhesive LST-R1 cells was observed and the adhesive cells were counted. Results Laser confocal scanning microscopy showed the expression of galectin-1 on the membrane of LST-R1 cells.The binding rate between galectin-1 and its antibody was 38.2%?0.92%. In the galectin-1 antibody inhibiting group,more and more irregular angular adhesive cells appeared, and most of the adhesive cells were growing in singles, while the adhesive cells,mainly clustered, in the control group were round or roudish. The numbers of adhesive LST-R1 cells in the antibody inhibiting group and the control group were 40 4 and 30 4 respectively (P

To present a data processing method of traditional Chinese medicine based on Apriori algorithm to improve the efficiency of data mining and ensure the accuracy of the knowledge or conclusion in the data mining. The importance of data preprocessing in data mining was analyzed, along with the characteristics of TCM data and the requirements of Apriori algorithm for mining data. Some new functions were formed with considerations on the exam-ples. The data preprocessing was explored from the aspects of terminology standardization, eliminating unqualified data, structured prescription data, data sorting and etc. The new functions were simple and easy to operate, and the preprocessed data made the efficiency of TCM data mining enhanced greatly. The preprocessing method based on Apriori algorithm for TCM data facilities the TCM data mining.

Objective To explore the genetic prenatal diagnosis method for acbendroplasia (ACH).Methods During May to November 2007, three ACH pedigrees were diagnosed at the Prenatal Diagnosis Center, Department of Obstetrics and Gynecology, Affiliated Drum Tower Hospital of Medical College, Nanjing University. In family 1, there was a 6-month-old male ACH infant. In family 2, the expectant mother, with 18 weeks of pregnancy, was an ACH patient. Amniocentesis was performed for prenatal diagnosis. The fetus of family 3 was diagnosed as ACH by ultrasound examination on the 39th week of gestation. Umbilical cord blood of this fetus was collected for examination. Totally, three methods, restriction enzyme (Sfc Ⅰ and Msp Ⅰ ) digestion analysis, denaturing high performance liquid chromatography (DHPLC) and sequencing analysis were performed simultaneously to detect the pathogenic mutation of flbroblastic growth factor receptor 3 (FGFR3) for the three ACH families. Results ( 1 ) The DHPLC detection: heteroduplex was detected in the patient of family 1 ; beth the patient and the fetus of family 2 showed heteroduplex results; the result of the fetus of family 3 was also heteroduplea. (2) The enzyme digestion analysis for the PCR products of 10 exon of FGFR3: after Sfc Ⅰ digestion, the PCR products of patients and the fetus of family 1 and 2 showed not only the band of 247 bp, but also bands of 162 bp and 85 bp. But their PCR products could not be digested by Msp Ⅰ , and it only showed the band of 247 bp. For the fetus of family 3, the PCR products could not be digested by Sfc Ⅰ , while after digestion by Msp Ⅰ , bands of 162 bp and 85 bp were shown up. The PCR products of the normal control could be digested by neither Sfc Ⅰ nor Msp Ⅰ. (3) The sequencing results: the heterozygote mutation of 1138 C→A was confirmed in the patient of family 1. The pregnant woman and her fetus in family 2 showed the same result. The heterozygote mutation of C→C was confirmed in the fetus of family 3. The site of 1138 was G homozygote in the normal control The three detection results of the fetus in family 2 were the same as that of the mother, which means that the fetus inherited the same pathogenic mutation from his or her mother. Conclusions Both DHPLC and restriction enzyme digestion analysis could detect the mutation of FGFR3 gene, but DHPLC is more rapid, convenient and sensitive. So DHPLC can be applied to genetic diagnosis and prenatal diagnosis for ACH patients.

Objective To construct the RNA interference (RNAi) eukaryotic expression vectors of galectin-1, and establish the LST-R1 cell lines stably transfected by RNAi vectors. Methods Two pairs of small interfering RNAs (siRNA) targeting to galectin-1 mRNA (GenBank: NM002305) were designed. The corresponding single-strand short hairpin interfering RNAs (shRNA), containing BamH Ⅰ, Hind Ⅲ sites and a 9nt hairpin structure, was synthesized and annealed. The annealed product and the linear eukaryotic expression plasmid pSuperior-puro, which were digested with Bgl Ⅱ and Hind Ⅲ, were ligated by T4 ligase to set up the interfering system. The recombined plasmid was identified with EcoR Ⅰand Hind Ⅲ enzyme digestion and sequencing, and co-transfected to LST-R1 cells with pcDNA6/TR with Lipofectamine2000. Positive clones were selected with 0.8?g/ml puromycin. After incubated with 4?g/ml tetracyclin for 48 hours, RT-PCR, Western blotting and immunochemistry were employed to determine galectin-1 mRNA and protein levels. Results Sequencing results suggested that the nucleotide sequence and read frame of RNAi eukaryotic exprssion vector of galectin-1, p-shRNA1 and p-shRNA2 were perfect. Stably transfected LST-R1 cell lines of p-shRNA1-LST and p-shRNA2-LST were established. The relative values of galectin-1 expression in LST-R1 cells, p-shRNA1-LST cells, p-shRNA2-LST cells and p-LST cells by RT-PCR were 0.616, 0.298, 0.373 and 0.641, respectively, and 1.00, 0.07, 0.38 and 0.97 by Western blotting. The p-shRNA1 gave the best interfering effect, which was in conformity with the results of immunochemistry measurement. Conclusions RNAi eukaryotic expression vector of galectin-1 mRNA has been successfully constructed. Establishment of the stably transfected LST-R1 cell lines may lay a foundation to explore the roles of galectin-1 in laterally spreading tumor.

Objective To observe the changes of LoVo cell growth by galectin-1.Methods Eukaryotic expression vector of galectin-1 was constructed and transfected into LoVo cells using lipofectamineRM 2000.Immunochemistry was employed to detect galectin-1 expresson.LoVo cell proliferation and apoptosis were observed.Results Galectin-1 eukaryotic expression vector pEGFP-C1/GAL1 was successfully constructed.Three cell clones,p-GAL1-LoVo and p-LoVo,transfected with pEGFP-C1/GAL1 and pEGFP-C1 correspondingly,and LoVo were cultured successfully.Galectin-1 expression downregulated Bcl-2 level only occurring in p-GAL1-LoVo cells.p-GAL1-LoVo cell proliferation was similar to p-LoVo and LoVo cells,but p-GAL1-LoVo cell apoptosis was increased(9.61?0.56)%,as compared with p-LoVo and LoVo cells,[(3.56?0.53)% and(3.46?0.46)% respectively](P

? carotene could decrease significantly the ear index and the quantity of Evan' s blue exudation that were skin photosensitive side actions in mice treated by photodynimic therapy ( PDT ) . The PGE 1 content in skin and the histamine content in skin and blood of mice were remarkably raised by raised by PDT. For example, the PGE 1 content in hematoporphyrin derivative group was 6.4 fold greater than that of control group. The PGE1 and histamine cotents were obviously decreased to nearly normal level by pretreatment with ? carotene. The results indicated that ? carotene can antagonise the skin photosensitive side action induced by PDT and the action was related to the decrease of PGE1 and histamine content in skin of mice. The cyclo-oxidase inhibitor indomethacin and the H1 receptor blocker benadryl could also decrease ear index and the quantity of Evan' s blue exudation

Objective To explore the endoscopic and histopathological features of serrated adenomas (SAs).Methods The data of patients with colorectal polyps diagnosed in the Digestive Endoscopy Center at Nanfang Hospital from January 2002 to July 2005 were reviewed and the detection rate, endoscopic appearances, pit patterns and histopathological features of SAs were analyzed.Results In 1928(16.21%) out of 11 894 patients undergoing colonscopy 2811 polyps were found.Among them 61 patients with 71 polyps were found,with a detection rate of 0.51%.The SAs,larger than hyperplastic polyps obviously,were found in patients 39.44% with diameter (larger than 1 cm).The incidence of pedunculated polyps in SAs (26.76%) was higher than that in hyperplastic polyps(13.25%),but less than in adenomatous polyps (43.95%).The pit patterns of SAs, were typeⅢ pit pattern (41.67%) and type Ⅳ pit pattern (18.33%), this result was similar to adenomatous polyps.The incidences of moderate and severe dysplasia of SAs were higher than those of tubular adenomas but lower than villous adenomas.The canceration rate of SA was 2.82%.Conclusion The endoscopic appearances,these of pit patterns and histopathological features of SAs,were different from hyperplastic polyps essentially, but similar to neoplastic polyps with potential malignancy,which should be emphasized in clinical practice.

Objective To investigate the application of "Guidelines for performing fetal cardiac scan", issued by the International Society of Ultrasound in Obstetries and Gynecology in 2006, in prenatal screening of fetal congenital heart disease (CHD). Method Totally, 5000 singleton pregnancies presented at the Maternal-Fetal Medical Center of the Affiliated Drum Tower Hospital of Nanjing University Medical School from September 2006 to July 2007, for prenatal screening were included in this study, with the median maternal age of 28 ( range, 18～48 ) and the median gestation of 27 ( range, 18～40 ) weeks. Ultrasound screenings were performed on each fetal heart according to "Guidelines for performing fetal cardiac scan" via the four-chamber and outflow tracts & three-vessel views and fetal echocardiographies were further conducted for suspected cases. Once congenital heart disease was confirmed, amniocentesis or cordocentesis was suggested for fetal karyotyping for ongoing pregnancies and autopsy was performed when the pregnancy was terminated after formal consent. Bom babies were followed up at 2～6 months of age using echocardiography. Result The four-chamber views were successfully obtained in 97.64% (4882/5000) of all the pregnancies , among which the left ventricular and right ventricular outflow tracts and three-vessel views were obtained in 87.69% ( 4281/4882 ), 82.51% ( 4028/4882 ) and 96.29% ( 4701/4882 ), respectively. Higher successful rate was found in the second trimester than the third trimester in obtaining the standard views (P<0.05). Finally, 73 (1.50%) among the 4882 cases were diagnosed as CHD. Fifty of them were diagnosed prenatally (24 cases in the second trimester and 26 cases in the third trimester) and 23 were missed and 1 misdiagnosed by prenatal ultrasound. Eighteen cases were found with extracardiac malformations. Autopsy was performed in 19 CHD which diagnosed prenatally, and all autopsy reports were consistent with ultrasound foundings. Twelve babies received postnatal echocardiography among which 11 were unanimous, and 1 baby diagnosed as tricuspid insufficiency prenatally was confirmed normal after birth. Abnormal karyotype was found in 7 out of the 23 who had karyotyping performed. Altogether, 28 cases were diagnosed by four chamber view only and 50 cases by combining other views, giving the sensitivity, specificity, false negative rate and false positive rate of 69% (50/73), 99.98% (4808/4809), 0.48% (23/4831) and 2% ( 1/51 ) ,respectively. Conclusion The "Guidelines for performing fetal cardiac scan" is practical and easy to abide by. The optimal time for fetal cardiac examination is at 18～27 weeks of gestation. Four-chamber view together with the outflow tracts and three-vessel views examination can detect 69% of CHD in utero.

OBJECTIVE:To study the genetic diversity of germplasm resources of Duchesnea indica from Hunan. METHODS:The random amplified polymorphic DNA(RAPD)reaction system of germplasm resources was established and RAPD-PCR was ad-opted to detect the expressions of 24 groups of D. indica from Hunan(including 21 wild and 3 cultivated varietos in different geo-graphical distribution). Agarose gel electrophoresis was used to analyze the results and calculate allele number (Na),effective al-lele number (Ne) the polymorphism points percentage (PPB),Nei’s gene diversity index (H) and Shannon’s information index (I). RESULTS:Totally 14 polymorphic primers were screened and 90 electrophoresis bands were amplified,including 81 polymor-phic bands with Na of 1.900 0,Ne of 1.540 1,PPB of 90.0%,H of 0.312 8 and I of 0.467 5. CONCLUSIONS:The genetic diver-sity of D. indica from Hunan is rich,and there were differences in genetic background between the cultivated species and wild spe-cies;clustering results show significant correlation with geographic distribution.